Journal of virological methods最新文献_第3页

Evaluation of the Alinity m CMV assay for detecting and quantifying cytomegalovirus DNA in non-plasma samples 评估用于检测和量化非血浆样本中巨细胞病毒 DNA 的 Alinity m CMV 检测法。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2024-11-19 DOI: 10.1016/j.jviromet.2024.115069

Anaïs Scohy , Maria A. Argudín , Florence Kabera , Nathalie Olive , Benoît Kabamba Mukadi

{"title":"Evaluation of the Alinity m CMV assay for detecting and quantifying cytomegalovirus DNA in non-plasma samples","authors":"Anaïs Scohy , Maria A. Argudín , Florence Kabera , Nathalie Olive , Benoît Kabamba Mukadi","doi":"10.1016/j.jviromet.2024.115069","DOIUrl":"10.1016/j.jviromet.2024.115069","url":null,"abstract":"<div><div>CMV infection remains a well-recognized threat in immunocompromised patients and is a major cause of congenital infection. If plasma and whole blood are routinely used as clinical samples for detection of CMV infection and disease, CMV laboratory testing in non-blood samples is becoming more and more relevant to support the diagnosis, prognosis and management of CMV infection. Accurate CMV viral load assessment in various body fluids is therefore essential. We evaluated the performance of the Alinity m CMV assay for detecting and quantifying CMV in plasma, amniotic fluid, bronchoalveolar lavage, cerebrospinal fluid, saliva and urine. Using a commercially available CMV reference panel and CMV culture supernatant, we assessed the linearity and accuracy of the Alinity m CMV assay across the different sample types. Excellent linear correlations (r values > 0.98) and a good accuracy (bias < ± 0.50 Log<sub>10</sub> IU/mL and SD < 0.23) were observed. In conclusion, the Alinity m CMV assay is suitable to detect and quantify CMV DNA in plasma but also in all non-plasma samples tested. Random and continuous access capabilities of the Alinity m enable rapid and efficient laboratory detection and quantification of CMV DNA.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"332 ","pages":"Article 115069"},"PeriodicalIF":2.2,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142687357","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Development of polyclonal chicken egg yolk immunoglobulin Y (IgY) antibodies targeting SARS-CoV-2 multi-epitope antigen 针对 SARS-CoV-2 多表位抗原的多克隆鸡卵黄免疫球蛋白 Y (IgY) 抗体的开发。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2024-11-17 DOI: 10.1016/j.jviromet.2024.115062

Azzania Fibriani, Katerina Naisanu, Nicholas Yamahoki, Denti Rizki Kinanti

{"title":"Development of polyclonal chicken egg yolk immunoglobulin Y (IgY) antibodies targeting SARS-CoV-2 multi-epitope antigen","authors":"Azzania Fibriani, Katerina Naisanu, Nicholas Yamahoki, Denti Rizki Kinanti","doi":"10.1016/j.jviromet.2024.115062","DOIUrl":"10.1016/j.jviromet.2024.115062","url":null,"abstract":"<div><div>Severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) is the primary cause of the Coronavirus disease 2019 (COVID-19) pandemic, which affects millions of people worldwide with high levels of infectivity and mortality. However, the antibodies developed for COVID-19 research and diagnostics are still limited. Therefore, in this study, we developed polyclonal immunoglobulin (IgY) antibodies from chicken egg yolk targeting multi-epitope antigen of SARS-CoV-2. After immunizing hens with a SARS-CoV-2 multi-epitope peptide, IgY antibodies were isolated from chicken eggs and further characterized using SDS-PAGE and ELISA. The results showed that the IgY antibodies were successfully isolated from egg yolks. The sandwich ELISA results demonstrated that the isolated IgYs could bind to SARS-CoV-2 antigens, both the multi-epitope peptide and the trimeric Spike. Furthermore, the developed polyclonal antibodies could recognize SARS-CoV-2 in human nasopharyngeal swab samples, even at the lowest concentration (dilution at 1:10000). Thus, it can be concluded that the developed polyclonal IgYs were successfully produced and have the potential to be applied in the development of COVID-19 diagnostics.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"331 ","pages":"Article 115062"},"PeriodicalIF":2.2,"publicationDate":"2024-11-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142647708","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Optimisation of a multiplexed, high throughput assay to measure neutralising antibodies against SARS-CoV-2 variants 优化多路复用高通量测定法，以测定针对 SARS-CoV-2 变体的中和抗体。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2024-11-16 DOI: 10.1016/j.jviromet.2024.115073

Caroline L. Ashley , Malik Bloul , Sibel Alca , Lachlan Smith , Wang Jin , David Khoury , Claudio Counoupas , Miles Davenport , James A. Triccas , Megan Steain

{"title":"Optimisation of a multiplexed, high throughput assay to measure neutralising antibodies against SARS-CoV-2 variants","authors":"Caroline L. Ashley , Malik Bloul , Sibel Alca , Lachlan Smith , Wang Jin , David Khoury , Claudio Counoupas , Miles Davenport , James A. Triccas , Megan Steain","doi":"10.1016/j.jviromet.2024.115073","DOIUrl":"10.1016/j.jviromet.2024.115073","url":null,"abstract":"<div><div>A multiplexed, lentivirus-based pseudovirus neutralisation assay (pVNT) was developed for high-throughput measurement of neutralising antibodies (nAbs) against three distinct SARS-CoV-2 spike variants. Intra-assay variability was minimised by optimising the plate layout and determining an optimal percentage transduction for the pseudovirus inoculum. Comparison of EC<sub>50</sub> titres between single and multiplexed pVNT assays showed no significant differences, indicating reliability of the multiplexed assay. Evaluation of convalescent human sera confirmed assay robustness, with consistent EC<sub>50</sub> titres for variant pseudoviruses relative to the ancestral strain observed across single and multiplexed assays. This multiplexed pVNT provides a reliable tool for assessing nAb responses against SARS-CoV-2 variants and could be used to accelerate preclinical vaccine assessment in preparation for the next coronavirus pandemic.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"332 ","pages":"Article 115073"},"PeriodicalIF":2.2,"publicationDate":"2024-11-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142668436","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Evaluation and comparison of three high throughput assays (Alinity m CMV, Aptima CMV Quant and cobas CMV) for quantifying CMV DNA in plasma samples 评估和比较用于定量血浆样本中 CMV DNA 的三种高通量检测方法（Alinity m CMV、Aptima CMV Quant 和 cobas CMV）。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2024-11-15 DOI: 10.1016/j.jviromet.2024.115068

Jodie D’Costa, Doris Chibo, Katherine Soloczynskyj, Mitchell Batty, Rizmina Sameer, Elaine Lee, Thomas Tran, Dimi Mavroulis, Megan Gooey, Eloise Williams, Kathy Jackson

{"title":"Evaluation and comparison of three high throughput assays (Alinity m CMV, Aptima CMV Quant and cobas CMV) for quantifying CMV DNA in plasma samples","authors":"Jodie D’Costa, Doris Chibo, Katherine Soloczynskyj, Mitchell Batty, Rizmina Sameer, Elaine Lee, Thomas Tran, Dimi Mavroulis, Megan Gooey, Eloise Williams, Kathy Jackson","doi":"10.1016/j.jviromet.2024.115068","DOIUrl":"10.1016/j.jviromet.2024.115068","url":null,"abstract":"<div><h3>Background</h3><div>Cytomegalovirus (CMV) can cause symptomatic CMV syndrome or tissue-invasive CMV disease in immunocompromised individuals, including solid-organ transplant and hematopoietic stem cell transplant recipients. In these populations, monitoring of CMV load is essential, assessing both risk of disease and response to antiviral therapy. High throughput commercial assays are currently available for CMV quantitation, but they are often evaluated independently, with few studies comparing these assays. This study evaluated CMV quantitative assays for use with the Roche cobas 6800, Abbott Alinity m and Hologic Panther platforms using stored patient plasma.</div></div><div><h3>Methods</h3><div>Analytical evaluation was performed using the 1st WHO international standard for human CMV for Nucleic Acid Amplification Techniques (cobas and Alinity m) or the Hologic CMV QC Calibrator 6 (Aptima). Parallel testing of 136 clinical plasma samples was performed across the three platforms.</div></div><div><h3>Results</h3><div>Linearity for each assay ranged from 98.6 % to 99.96 % and precision and limit of quantitation were as expected with little variation between platforms. 136 clinical plasma samples were evaluated with similar agreement observed between each assay. The greatest positive agreement was between the Aptima Quant and Alinity m assays (95.6 %, 95 % CI 89–98.6 %) and the lowest between the Aptima Quant and cobas assays (94.1 %, 87.4–97.5 %).</div></div><div><h3>Conclusions</h3><div>All assays were sensitive and accurate when quantifying CMV, and performance across all 3 assays was comparable for monitoring CMV viral loads in patient plasma.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"332 ","pages":"Article 115068"},"PeriodicalIF":2.2,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142647720","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Quantitative and qualitative analysis of seroconversion after one year of vaccination with inactivated SARS-CoV-2 vaccine (CoronaVac®) in healthcare workers: Cross-sectional analytical study "医护人员接种 SARS-CoV-2 灭活疫苗 (CoronaVac®) 一年后血清转换的定量和定性分析：横断面分析研究"。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2024-11-15 DOI: 10.1016/j.jviromet.2024.115067

Júlia Gomes da Silva , Viviana Galimberti Arruk , Glaucia Raquel Luciano da Veiga , Luiz Vinícius de Alcantara Sousa , Beatriz da Costa Aguiar Alves , Fernando Luiz Affonso Fonseca , Inneke Marie van der Heijden Natário

{"title":"Quantitative and qualitative analysis of seroconversion after one year of vaccination with inactivated SARS-CoV-2 vaccine (CoronaVac®) in healthcare workers: Cross-sectional analytical study","authors":"Júlia Gomes da Silva , Viviana Galimberti Arruk , Glaucia Raquel Luciano da Veiga , Luiz Vinícius de Alcantara Sousa , Beatriz da Costa Aguiar Alves , Fernando Luiz Affonso Fonseca , Inneke Marie van der Heijden Natário","doi":"10.1016/j.jviromet.2024.115067","DOIUrl":"10.1016/j.jviromet.2024.115067","url":null,"abstract":"<div><div>A descriptive study was carried out with health professionals in Sao Paulo city. Were included individuals vaccinated with 2 doses of the inactivated vaccine. Demographic, clinical and vaccination information was obtained from professionals with or without comorbidities. Two serological assays were used to identify the presence and quantity of anti-Spike IgG in serum samples. 433 healthy healthcare professionals were included and 58.9 % completed the 4 clinical stages of serological assessment. Among adults and elderly people, 25.2 % had chronic diseases (hypertension 50.5 %, diabetes 10 % and obesity 6.5 %). Most individuals have 95 % protection in the first 3 months after the second dose, and 67.68 % protection after 6 months. Total antibodies ranged from 3 to 10 on the reactivity index, and the anti-RBD IgG levels were high. CoronaVac has a 94 % seroconversion rate after 2 doses and can prevent serious cases and outbreaks of the disease, if used on a large scale.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"332 ","pages":"Article 115067"},"PeriodicalIF":2.2,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142647543","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Mapping and quantification of potato virus A RNA genomes within viral particles and polysomes in infected plant cells 绘制受感染植物细胞中病毒颗粒和多聚体内马铃薯病毒 A RNA 基因组的图谱并对其进行定量。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2024-11-15 DOI: 10.1016/j.jviromet.2024.115066

Pinky Dutta, Kristiina Mäkinen

{"title":"Mapping and quantification of potato virus A RNA genomes within viral particles and polysomes in infected plant cells","authors":"Pinky Dutta, Kristiina Mäkinen","doi":"10.1016/j.jviromet.2024.115066","DOIUrl":"10.1016/j.jviromet.2024.115066","url":null,"abstract":"<div><div>Potato virus A belongs to the genus <em>Potyvirus,</em> a group of single-stranded positive sense RNA viruses infecting crops worldwide. To initiate infection in a host, its genome takes part in different activities, <em>viz.</em>, translation, replication, encapsidation during the infection cycle. Extensive research has been carried out to scrutinize the stages of potyviral infection cycle and decipher the strategies it employs to cause disease. Nonetheless, the amount of viral RNA taking part in translation and virion formation, at a given time point, is missing. In this study, we quantified the percentage of viral RNA that exists as virions and those that associates with host polysome, relative to total viral RNA in infected plant tissue. We employed a revised version of immuno-capture reverse transcription PCR and polysome profiling to address our queries. We tested three different coating antibody concentrations and further optimized the immuno-capture reverse transcription PCR protocol to address its limitation of binding and retaining viral particles. Our results indicate that most of the viral RNA (69 %) exists as encapsidated genomes, while 3 % of total viral RNA associates with host polysomes. These findings are crucial for correct interpretation of quantitative translational studies in which correlation must be made between the number of polysome-associated transcripts and the amount of protein synthesized.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"332 ","pages":"Article 115066"},"PeriodicalIF":2.2,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142644280","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Wastewater sample storage for physicochemical and microbiological analysis 用于理化和微生物分析的废水样本存储。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2024-11-14 DOI: 10.1016/j.jviromet.2024.115063

Gordon Webster , Shrinivas Nivrutti Dighe , William B. Perry , Ewan H. Stenhouse , Davey L. Jones , Peter Kille , Andrew J. Weightman

{"title":"Wastewater sample storage for physicochemical and microbiological analysis","authors":"Gordon Webster , Shrinivas Nivrutti Dighe , William B. Perry , Ewan H. Stenhouse , Davey L. Jones , Peter Kille , Andrew J. Weightman","doi":"10.1016/j.jviromet.2024.115063","DOIUrl":"10.1016/j.jviromet.2024.115063","url":null,"abstract":"<div><div>Wastewater-based epidemiology (WBE) is a crucial tool for health and environmental monitoring, providing real-time data on public health indicators by analysis of sewage samples. Ensuring the integrity of these samples from collection to analysis is paramount. This study investigates the effects of different cold-storage conditions on the integrity of wastewater samples, focusing on both microbiological markers (such as extractable nucleic acids, SARS-CoV-2, and crAssphage) and physicochemical parameters (including ammonium, orthophosphate, pH, conductivity, and turbidity). Composite samples from the raw wastewater influent from five wastewater treatment works in South Wales, UK, were stored at 4°C, -20°C, and -80°C, and subjected to up to six freeze-thaw cycles over one year. The study found significant effects of storage temperature on the preservation of certain WBE markers, with the best yield most frequently seen in samples stored at -80°C. However, the majority of WBE markers showed no significant difference between storage at -80°C or at 4°C, demonstrating that it may not always be necessary to archive wastewater samples at ultra-low temperatures, thus reducing CO<sub>2</sub> emissions and laboratory energy costs. These findings underscore the importance of optimized storage conditions to maintain sample integrity, while ensuring accurate and reliable WBE data for public health and environmental monitoring.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"332 ","pages":"Article 115063"},"PeriodicalIF":2.2,"publicationDate":"2024-11-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142639191","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Evaluation of the impact of hollow fiber pore size and membrane material on virus concentration using a handheld hollow fiber method 使用手持式中空纤维法评估中空纤维孔径和膜材料对病毒浓度的影响。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2024-11-14 DOI: 10.1016/j.jviromet.2024.115065

Shinsuke Higuchi , Tatsuki Satou , Yuki Uchida

引用次数: 0

Validation of a molecular multiplex assay for the simultaneous detection and differentiation of bluetongue virus and epizootic haemorrhagic disease virus in biological samples 验证同时检测和区分生物样本中蓝舌病病毒和流行性出血病病毒的分子多重分析法。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2024-11-13 DOI: 10.1016/j.jviromet.2024.115064

Ottavio Portanti, Eugenia Ciarrocchi, Roberta Irelli, Andrea Palombieri, Romolo Salini, Irene Melegari, Maura Pisciella, Simone Pulsoni, Daria Di Sabatino, Massimo Spedicato, Giovanni Savini, Alessio Lorusso

{"title":"Validation of a molecular multiplex assay for the simultaneous detection and differentiation of bluetongue virus and epizootic haemorrhagic disease virus in biological samples","authors":"Ottavio Portanti, Eugenia Ciarrocchi, Roberta Irelli, Andrea Palombieri, Romolo Salini, Irene Melegari, Maura Pisciella, Simone Pulsoni, Daria Di Sabatino, Massimo Spedicato, Giovanni Savini, Alessio Lorusso","doi":"10.1016/j.jviromet.2024.115064","DOIUrl":"10.1016/j.jviromet.2024.115064","url":null,"abstract":"<div><div>Bluetongue virus (BTV) and epizootic haemorrhagic disease virus (EHDV) are <em>Culicoides</em>-transmitted viruses, circulating in multiple serotypes, that cause two relevant WOAH-listed diseases of ruminants. Following its first identification in Tunisia in 2021, a novel EHDV strain belonging to serotype 8 has been detected in cattle showing BTV-like symptoms in Italy and Spain in 2022, and soon after in Portugal and France. These are European regions with recurrent circulations of different BTV serotypes. Hence, in this study we describe the validation of a TaqMan RT-qPCR panBTV/panEHDV assay, based on well-established primers and probes sets, able to simultaneously detect and distinguish between BTV and EHDV. The implemented assay, characterized by high sensitivity, specificity and good reproducibility, can be successfully applied for the rapid and affordable diagnosis needed in the current epidemiological situation and represents a powerful tool to be employed in surveillance and control strategies with a significant reduction of costs.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"332 ","pages":"Article 115064"},"PeriodicalIF":2.2,"publicationDate":"2024-11-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142622902","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Design, development, and validation of a strand-specific RT-qPCR assay to differentiate replicating versus nonreplicating Rabies virus 设计、开发和验证用于区分复制与非复制狂犬病病毒的链特异性 RT-qPCR 检测方法。

IF 2.2 4区医学

Journal of virological methods Pub Date : 2024-11-08 DOI: 10.1016/j.jviromet.2024.115054

Hong Xiang, Chao Hong, Yuan Tian, Yu Gao, Yina Cun, Hongling Zhao, Guilan Li, Yu Chen, Jian Zhou

{"title":"Design, development, and validation of a strand-specific RT-qPCR assay to differentiate replicating versus nonreplicating Rabies virus","authors":"Hong Xiang, Chao Hong, Yuan Tian, Yu Gao, Yina Cun, Hongling Zhao, Guilan Li, Yu Chen, Jian Zhou","doi":"10.1016/j.jviromet.2024.115054","DOIUrl":"10.1016/j.jviromet.2024.115054","url":null,"abstract":"<div><div>The <em>Rabies virus</em>, a single-strand RNA virus with a negative-sense polarity, is responsible for causing encephalitis and is a zoonotic disease. If not promptly treated after infection, it has a close to 100 % fatality rate. Similar to other negative-sense polarity single-strand RNA viruses, the <em>Rabies virus</em> requires the creation of a positive-strand RNA intermediate for replication. One approach to identify this replication activity is to detect the complementary strand of the viral RNA genome in suspected infected cells or tissues. The reported <em>Rabies virus</em> RT-qPCR detection methods are designed to detect total viral load in samples without distinguishing between positive- and negative-strand for RNA viruses. As such, in this study, a sensitive Taqman-based strand-specific RT-qPCR assay has been developed to quantitatively detect both the positive- and negative-strand of the <em>Rabies virus</em>. This method demonstrates good reproducibility across a wide dynamic range and exhibits linearity of 8 logs with a lower limit of detection of 10<sup>3</sup> copies/μL for the positive-strand and 9 logs with a lower limit of detection of 10<sup>2</sup> copies/μL for the negative-strand. Notably, it can accurately detect a specific viral RNA strand even in the presence of high levels of the opposite strand, confirming the method’s specificity. In summary, a reliable strand-specific RT-qPCR assay has been developed and validated to differentiate replicating from non-replicating <em>Rabies virus</em>.</div></div>","PeriodicalId":17663,"journal":{"name":"Journal of virological methods","volume":"332 ","pages":"Article 115054"},"PeriodicalIF":2.2,"publicationDate":"2024-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142622759","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0